Dataset Information

Gga-miR-449b-5p Regulates Steroid Hormone Synthesis in Laying Hen Ovarian Granulosa Cells by Targeting the IGF2BP3 Gene.

ABSTRACT: MiRNAs have been found to be involved in the regulation of ovarian function as important post-transcriptional regulators, including regulators of follicular development, steroidogenesis, cell atresia, and even the development of ovarian cancer. In this study, we evaluated the regulatory role of gga-miR-449b-5p in follicular growth and steroid synthesis in ovarian granulosa cells (GCs) of laying hens through qRT-PCR, ELISAs, western blotting and dual-luciferase reporter assays, which have been described in our previous study. We demonstrated that gga-miR-449b-5p was widely expressed in granulosa and theca layers of the different-sized follicles, especially in the granulosa layer. The gga-miR-449b-5p had no significant effect on the proliferation of GCs, but could significantly regulate the expression of key steroidogenesis-related genes (StAR and CYP19A1) (p < 0.01) and the secretion of P4 and E2 (p < 0.01 and p < 0.05). Further research showed that gga-miR-449b-5p could target IGF2BP3 and downregulate the mRNA and protein expression of IGF2BP3 (p < 0.05). Therefore, this study suggests that gga-miR-449b-5p is a potent regulator of the synthesis of steroid hormones in GCs by targeting the expression of IGF2BP3 and may contribute to a better understanding of the role of functional miRNAs in laying hen ovarian development.

SUBMITTER: Wu X

PROVIDER: S-EPMC9559480 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature

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gga-miR-449b-5p Regulates Steroid Hormone Synthesis in Laying Hen Ovarian Granulosa Cells by Targeting the IGF2BP3 Gene.

Wu Xing X Zhang Na N Li Jing J Zhang Zihao Z Guo Yulong Y Li Donghua D Zhang Yanhua Y Gong Yujie Y Jiang Ruirui R Li Hong H Li Guoxi G Liu Xiaojun X Kang Xiangtao X Tian Yadong Y

Animals : an open access journal from MDPI 20221009 19

MiRNAs have been found to be involved in the regulation of ovarian function as important post-transcriptional regulators, including regulators of follicular development, steroidogenesis, cell atresia, and even the development of ovarian cancer. In this study, we evaluated the regulatory role of gga-miR-449b-5p in follicular growth and steroid synthesis in ovarian granulosa cells (GCs) of laying hens through qRT-PCR, ELISAs, western blotting and dual-luciferase reporter assays, which have been de ...[more]

PMID: 36230451

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Project description:MiRNAs are typically reported to play a negative regulatory role in post-transcriptional expression of target genes and are widely involved in a variety of biological processes such as growth, metabolism and reproduction. However, research on the role of miRNAs in the ovulation process of chicken ovaries is still insufficient compared to that in mammals. Here, we investigated the regulatory mechanisms of gga-miR-6634-5p in the growth and steroid hormone secretion of chicken granulosa cells (GCs) by targeting MMP16. We found that gga-miR-6634-5p significantly down-regulated the mRNA levels of proliferation-related genes (CCND1, CDK1, and CDK6), decreased cell viability, the number of EdU-labelled positive cells, and the percentage of S-phase cells, as analysed by quantitative real time PCR (qRT-PCR), cell counting kit-8 (CCK-8), 5-ethynyl-2'deoxyuridine (EdU) and flow cytometry analyses (P < 0.01 or P < 0.05). qRT-PCR and enzyme-linked immunosorbent assay (ELISA) results demonstrated that gga-miR-6634-5p up-regulated the expression of steroid synthesis-related genes (CYP19A1, 3β-HSD, StAR and FSHR) (P < 0.01 or P < 0.05), as well as the secretion of estradiol (E2) and progesterone (P4) (P < 0.01 or P < 0.05). Furthermore, we found that MMP16 protein and gene expression can be down-regulated by gga-miR-6634-5p and demonstrated that MMP16 is a target gene of gga-miR-6634-5p by dual luciferase reporter assay (P < 0.05). In addition, we found that MMP16 stimulated the proliferation of GCs, significantly inhibited the expression of steroid synthesis related genes (CYP19A1, StAR, 3β-HSD and FSHR), and decreased the secretion of E2 and P4 (P < 0.01 or P < 0.05), which was consistent with the inhibitory effect of gga-miR-6634-5p. It was further found by functional enrichment analysis, qRT-PCR, western blot (WB) and ELISA that MMP16 may play a regulatory role in GCs proliferation as well as steroid hormone secretion through the mTOR signaling pathway and PPAR signaling pathway. Therefore, this study demonstrates that gga-miR-6634-5p modulates the proliferation of chicken GCs and the secretion of steroid hormones by targeting MMP16, which may contribute to a better understanding of the functional mechanisms of miRNAs in the ovarian development of laying hens.

Dataset Information

Gga-miR-449b-5p Regulates Steroid Hormone Synthesis in Laying Hen Ovarian Granulosa Cells by Targeting the IGF2BP3 Gene.

Publications

gga-miR-449b-5p Regulates Steroid Hormone Synthesis in Laying Hen Ovarian Granulosa Cells by Targeting the IGF2BP3 Gene.

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